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中华实验和临床感染病杂志(电子版) ›› 2020, Vol. 14 ›› Issue (02) : 110 -116. doi: 10.3877/cma.j.issn.1674-1358.2020.02.005

所属专题: 经典病例 经典病例 文献

论著

五例恙虫病东方体感染基因型及其基因变异
姚立农1, 陆群英2,(), 阮卫1, 占喆2, 朱水荣2, 姜理平2   
  1. 1. 310051 杭州市,浙江省疾病预防控制中心传染病防治所
    2. 310051 杭州市,浙江省疾病预防控制中心微生物所
  • 收稿日期:2019-06-11 出版日期:2020-04-15
  • 通信作者: 陆群英
  • 基金资助:
    2018年浙江省卫生科技项目(No. 2018KY340)

Gene genotypes and variation of 5 cases with Orientia tsutsugamushi infection

Linong Yao1, Qunying Lu2,(), Wei Ruan1, Zhe Zhan2, Shuirong Zhu2, Liping Jiang2   

  1. 1. Department of Communicable Diseases Control and Prevention, Zhejiang Province Center for Diseases Control and Prevention, Hangzhou 310051, China
    2. Department of Microbiology, Zhejiang Province Center for Diseases Control and Prevention, Hangzhou 310051, China
  • Received:2019-06-11 Published:2020-04-15
  • Corresponding author: Qunying Lu
  • About author:
    Corresponding author: Lu Qunying, Email:
引用本文:

姚立农, 陆群英, 阮卫, 占喆, 朱水荣, 姜理平. 五例恙虫病东方体感染基因型及其基因变异[J/OL]. 中华实验和临床感染病杂志(电子版), 2020, 14(02): 110-116.

Linong Yao, Qunying Lu, Wei Ruan, Zhe Zhan, Shuirong Zhu, Liping Jiang. Gene genotypes and variation of 5 cases with Orientia tsutsugamushi infection[J/OL]. Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition), 2020, 14(02): 110-116.

目的

旨在了解浙闽地区人群恙虫病东方体感染的基因型,并分析其基因变异。

方法

从浙闽地区5例散发恙虫病发热期患者血中分离病原体并进行细胞培养;提取感染细胞DNA,巢式PCR扩增完整恙虫病东方体56-kD型特异性抗原(TSA)基因和热休克蛋白基因(groESL)并测序;采用MEGA 7.0软件,进行序列比对和系统发育分析。

结果

病原学确认5例恙虫病患者,并分离到恙虫病东方体菌株。序列比对表明,5菌株中有2株的56-kD TSA基因和groESL基因100%一致,另2菌株的此二个基因序列一致性亦为100%,分别将两组菌暂时命名为浙江-1型和浙江-2型。56-kDa TSA基因比对和系统发育分析表明,浙江-1型和浙江-2型分别与台湾-A基因型和Gilliam-C基因型亲缘较近(98.45%和98.50%),但有明显变异;另1株菌Wuj/2014与台湾-A基因型高度相似(99.94%)。56-kDa TSA基因各基因型支系的时间树分析表明,台湾-A基因型、浙江-1型和浙江-2型3支系与祖先的分歧时间相对其他原型株晚,尤其是浙江-2型,说明这些基因型或亚型在恙虫病东方体的进化过程中出现较晚。

结论

本研究病例所感染恙虫病东方体基因型不同,可能为未被认识的新的基因亚型,迫切需要进行全基因组测序确认,并探讨其基因变异与人恙虫病严重程度间的关系。

Objective

To investigate the genotype of Orientia tsutsugamushi (O. tsutsugamushi) infection in Zhejiang and Fujian provinces, southeastern China and to analyze the gene variation.

Methods

Strains of O. tsutsugamushi were isolated from the blood of 5 scrub typhus patients of febrile phase and then cell culture were carried out. DNA from the infected cells were extracted and full-length 56-kD type-specific antigen (TSA) gene and full-length heat shock protein gene (groESL) were amplified. The gene products were sequenced. Multiple sequence alignment and phylogenetic analyses were conducted by Mega 7.0 software.

Results

Five febrile patients with O. tsutsugamushi infection were confirmed etiologically and five O. tsutsugamushi strains were isolated. The sequence alignment of 56-kD TSA gene and groESL gene showed that 2 among the 5 strains had 100% identity; while other two strains also had 100% identity of these two genes. These two groups were temporarily named Zhejiang-1 and Zhejiang-2. The 56-kDa TSA gene alignment and phylogenetic analysis showed that Zhejiang-1 was closely related to Taiwan-A genotype (98.45%) and Zhejiang-2 was closely related to Gilliam-C genotype (98.50%), but there were obvious variations. The other 1 strain of Wuj/2014 was significantly similar to Taiwan-A genotype (99.94%). Based on 56-kDa TSA gene, the timetree based on the RelTime method showed that Taiwan-A genotype, Zhejiang-1 and especially Zhejiang-2 genotype were later than other prototypes on the divergence time from their ancestors of each genotypic clade. These genotypes or subtypes emerged later in evolutionary process of O. tsutsugamushi.

Conclusions

There may be unrecognized novel O. tsutsugamushi variants among the patients in this study. The relationship between genetic variation and severity of scrub typhus also should be discussed.

表1 56-kDaTSA基因和groESL基因巢式PCR引物
图1 浙江-1型和浙江-2型56 kDa TSA蛋白VDⅠ和VDⅡ氨基酸变异
图2 基于56-kDaTSA基因构建的系统发育树
表2 本研究分离菌株与参考菌株56-kDa TSA基因和groESL基因序列的一致性
图3 基于GroESL基因构建的系统发育树(ML)
图4 基于RelTime时间树ML法估计的分歧时间
[1]
Tamura A, Ohashi N, Urakami H, et al. Classification of Rickettsia tsutsugamushi in a new genus, Orientia gen. nov., as Orientia tsutsugamushi comb. nov[J]. Int J Syst Bacteriol,1995,45(3):589-591.
[2]
Walker DH. Scrub Typhus-scientific neglect, ever-widening impact[J]. N Engl J Med,2016,375(10):913-915.
[3]
Luce-Fedrow A, Lehman ML, Kelly DJ, et al. A review of scrub typhus (Orientia tsutsugamushi and related organisms): then, now, and tomorrow[J]. Trop Med Infect Dis,2018,3(1):E8.
[4]
Bonell A, Lubell Y, Newton PN, et al. Estimating the burden of scrub typhus: A systematic review[J]. PLoS Negl Trop Dis,2017,11(9):e0005838.
[5]
Yang YF, Wang JL, Yao YC. A survey on scrub typhus found first time in Shandong Province[J]. Chin J Epidemiol,1987,8(5):280-284.
[6]
Zhang M, Zhao ZT, Wang XJ, et al. Genetic variants of Orientia tsutsugamushi in domestic rodents, Northern China[J]. Emerg Infect Dis,2013,19(7):1135-1137.
[7]
Hu J, Tan Z, Ren D, et al. Clinical characteristics and risk factors of an outbreak with scrub typhus in previously unrecognized areas, Jiangsu Province, China 2013[J]. PLoS One,2015,10(5):e0125999.
[8]
Cao M, Che L, Zhang J, et al. Determination of scrub typhus suggests a new epidemic focus in the Anhui Province of China[J]. Sci Rep,2016,6:20737.
[9]
Sun Y, Wei YH, Yang Y, et al. Rapid increase of scrub typhus incidence in Guangzhou, southern China, 2006-2014[J]. BMC Infect Dis,2017,17(1):13.
[10]
Li W, Dou X, Zhang L, et al. Laboratory diagnosis and genotype identification of scrub typhus from Pinggu district, Beijing, 2008 and 2010[J]. Am J Trop Med Hyg,2013,89(1):123-129.
[11]
Zheng L, Yang HL, Bi ZW, et al. Epidemic characteristics and spatio-temporal patterns of scrub typhus during 2006-2013 in Tai’an, Northern China[J]. Epidemiol Infect,2015,143(11):2451-2458.
[12]
Wu YC, Qian Q, Soares Magalhaes RJ, et al. Spatiotemporal dynamics of scrub typhus transmission in mainland China, 2006-2014[J]. PLoS Negl Trop Dis,2016,10(8):e0004875.
[13]
Xin HL, Yu JX, Hu MG, et al. Evaluation of scrub typhus diagnosis in China: analysis of nationwide surveillance data from 2006 to 2016[J]. Infect Dis Poverty,2019,8(1):59.
[14]
中国疾病预防控制中心传染病所立克次体研究室. 立克次体病诊断与处置手册[M]. 北京: 卫生部办公厅,2008.
[15]
Lu Q, Yu J, Yu L, et al. Rickettsia japonica Infections in Humans, Zhejiang Province, China, 2015[J]. Emerg Infect Dis,2018,24(11):2077-2079.
[16]
Yang HH, Huang IT, Lin CH, et al. New genotypes of Orientia tsutsugamushi isolated from humans in eastern Taiwan[J]. PLoS One,2012,7(10):e46997.
[17]
Lin PR, Tsai HP, Tsui PY, et al. Genetic typing, based on the 56-kilodalton type-specific antigen gene, of Orientia tsutsugamushi strains isolated from chiggers collected from wild-caught rodents in Taiwan[J]. Appl Environ Microbiol,2011,77(10):3398-3405.
[18]
Enatsu T, Urakami H, Tamura A. Phylogenetic analysis of Orientia tsutsugamushi strains based on the sequence homologies of 56-kDa type-specific antigen genes[J]. FEMS Microbiol Lett,1999,180(2):163-169.
[19]
Jeong HW, Choi YK, Baek YH, et al. Phylogenetic analysis of the 56-kDa type-specific protein genes of Orientia tsutsugamushi in Central Korea[J]. J Korean Med Sci,2012,27(11):1315-1319.
[20]
Ruang-Areerate T, Jeamwattanalert P, Rodkvamtook W, et al. Genotype diversity and distribution of Orientia tsutsugamushi causing scrub typhus in Thailand[J]. J Clin Microbiol,2011,49(7):2584-2589.
[21]
Duong V, Mai TT, Blasdell K, et al. Molecular epidemiology of Orientia tsutsugamushi in Cambodia and Central Vietnam reveals a broad region-wide genetic diversity [J]. Infect Genet Evol,2013,15(1):35-42.
[22]
Zhang L, Bi Z, Kou Z, et al. Scrub typhus caused by Orientia tsutsugamushi Kawasaki-related genotypes in Shandong Province, Northern China [J]. Infect Genet Evol,2015,30:238-243.
[23]
De W, Jing K, Huan Z, et al. Scrub typhus, a disease with increasing threat in Guangdong, China[J]. PLoS One,2015,10(2):e0113968.
[24]
张倩,刘运喜,吴晓明, 等. 中国内蒙古,新疆部分地区鼠类自然感染恙虫病东方体的调查[J]. 中华流行病学杂志,2006,27(6):475-478.
[25]
任立松,党荣理,马德新, 等. 新疆部分地区人群及动物感染恙虫病东方体调查[J]. 热带医学杂志,2012,12(10):1249-1257.
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